Subspecies Concept Research Articles

Genomic Data Clarify Aquarana Systematics and Reveal Isolation-by-Distance Dominates Phylogeography of the Wide-Ranging Frog Rana clamitans

Wide-ranging species often span geographic dispersal barriers, providing opportunity for divergence via genetic drift or natural selection. Such conditions can be favorable for speciation, and wide-ranging taxa are frequently subdivided into multiple species by modern molecular studies. However, with wide-ranging species, it is important to explicitly test for isolation-by-distance (IBD), which can produce continuous genetic variation that may be misinterpreted as discrete population structure or even distinct species. Here we examine the Green Frog (Rana clamitans), a wide-ranging species of the Aquarana clade distributed across much of North America. The broader phylogenetic context for R. clamitans within Aquarana is poorly understood, particularly its relationship with the closely related and IUCN Vulnerable Florida Bog Frog (Rana okaloosae). Additionally, although phenotypic variation within Rana clamitans previously merited description of two subspecies, patterns of genetic diversity remain unclear. Using genome-wide ddRAD markers, we largely resolve relationships within Aquarana and unambiguously identify R. okaloosae as the sister lineage to R. clamitans. Despite mtDNA introgression, we find little genomic evidence of hybridization between R. okaloosae and R. clamitans. Within R. clamitans there are two well-supported and geographically divided clades, which are identified as distinct species by a multispecies coalescent-based approach. However, the two clades exhibit low genealogical divergence. Population genetic analyses reveal that genetic variation within R. clamitans is best described by a pattern of IBD rather than independently evolving lineages. We conclude that R. clamitans is indeed a single species and the subspecies concept is of limited use. Our analysis demonstrates the importance of understanding continuous genetic variation when delimiting lineages and highlights the power of combining population genetic and phylogenetic perspectives to describe diversity within wide-ranging taxa.

A slippery reality: the epistemological shifting sands of tokogeny, phylogeny, lineages and species delimitation

This essay provides an alternative perspective on the history of classification, species and subspecies concepts to that presented in a recent review by Padial and de la Riva. Central points of difference relate to Darwin’s role in revolutionizing classification, Willi Hennig’s role in the development of the phylogenetic species concept, the metaphysics and epistemology of species as lineage segments, the true history of the subspecies concept, and the impact of large amounts of genetic data on the accuracy and precision of species/subspecies delimitation. Possession of a clear understanding of the history of systematics, and a chronology of which ideas inform which other ideas, provides critical background knowledge to build a sound conceptual framework for interpreting the nuances of phylogenetic evidence today. A glossary of potentially unfamiliar terms is included as an appendix.

Alpha-taxonomy in the cricetid rodent Neomicroxus, a first assessment

Neomicroxus , a recently named genus, comprises small-bodied cricetid rodents patchily distributed in high-Andean ranges from Ecuador to Venezuela. Currently, two species of Neomicroxus are recognized, N. bogotensis , endemic to the Cordillera Oriental in Colombia and Cordillera de Merida and Paramo de Tama in Venezuela, and N. latebricola that occurs northern Andes of Ecuador. The genus is among the most poorly understood Neotropical rodents and to date no formal assessment about its alpha taxonomy was conducted. Based on DNA evidence of the first portion of the mitochondrial cytochrome b gene (cytb) and the first exon of the interphotoreceptor retinoid binding protein (IRBP), as well as craniodental measurements, we explored the divergence degree, genetic structure and phyletic relationships of the two species currently allocated under Neomicroxus . Our analyses support the monophyly of the genus as well as its uncertain tribal affiliation. Neomicroxus was retrieved as structured in two main branches, in agreement with the traditional recognition of two species. The populations referred to N. bogotensis exhibit deep divergence values (> 6 %) pointing to the existence of undescribed species under its concept. In contrast, populations of N. latebricola show a shallow genetic structure although implying recognizable geographical breaks. A moderate degree of genetic and morphological differentiation supports a new subspecies for the western populations of N. latebricola . Our contribution is the first attempt to better understanding the alpha taxonomy of Neomicroxus , highlighting the importance of the geographic complexity as a barrier to the genetic flow in N. bogotensis and the significance of the subspecies concept to formalize the geographic variation recovered in N. latebricola

Are subspecies (ofEira barbara) real?

AbstractThe subspecies concept is one of the most controversial in Linnean taxonomy. In the past, subspecies were described without a clear conceptual framework, triggering confusion and motivating criticism of the very concept of a subspecies. At present, subspecies are conceived as aggregates of populations that are geographically isolated, are composed of interfertile individuals, and are morphologically diagnosable. The tayra, Eira barbara, was described in 1758 and has had a stable taxonomic history at the species level. However, below the species level, 16 subspecies have been named, with from two to seven subspecies recognized as valid by different authors. None of the subspecies were, however, described within a clear conceptual framework. Using the modern concept of a subspecies, I analyzed subspecies of E. barbara recognized by recent authors. I gathered morphometric data from 155 specimens in mammal collections, georeferenced each specimen, and recorded membership to subspecies assigned by different references and by its location. I gathered climate and geographic data for each location. I analyzed data using Principal Components Analysis (PCA) and analysis of variance (ANOVA). Specimens exhibited sexual dimorphism in size but not in skull shape. I used regression analysis to test for associations between skull shape and size and climate data. Geographic analyses documented that subspecies are not allopatric, violating one of the main properties of the subspecies concept. ANOVA showed significant differences in skull morphology between some pairs of recognized subspecies but not others. However, none of the subspecies segregated in the PCA. Thus, the recognized subspecies could not be diagnosed from morphological data, violating another property of the subspecies concept. Size varied greatly between the sexes using different schemes for recognized subspecies. Climate variables explained between 4% and 6% of size variation for males and females. Skull shape proved not to be geographically variable.

Shining a light on species delimitation in the tree genus Engelhardia Leschenault ex Blume (Juglandaceae)

Enhanced efficacy in species delimitation is critically important in biology given the pending biodiversity crisis under global warming and anthropogenic activity. In particular, delineation of traditional classifications in view of the complexity of species requires an integrative approach to effectively define species boundaries, and this is a major focus of systematic biology. Here, we explored species delimitation of Engelhardia in tropical and subtropical Asia. In total, 716 individuals in 71 populations were genotyped using five chloroplast regions, one nuclear DNA region (nrITS), and 11 nuclear simple sequence repeats (nSSR). Phylogenetic trees were constructed and relationships among species were assessed. Molecular analyses were then combined with 14 morphological characteristics of 720 specimens to further explore the species boundaries of Engelhardia. Integrating phylogenetic and morphological clusters provided well-resolved relationships to delineate seven species. The results suggested that: first, that E. fenzelii, E. roxburghiana, E. hainanensis, E. apoensis, and E. serrata are distinct species; second, E. spicata var. spicata, E. spicata var. aceriflora, E. spicata var. colebrookeana, and E. rigida should be combined under E. spicata and treated as a species complex; third, E. serrata var. cambodica should be raised to species level and named E. villosa. We illuminated that bias thresholds determining the cluster number for delimiting species boundaries were substantially reduced when morphological data were incorporated. Our results urge caution when using the concepts of subspecies and varieties in order to prevent confusion, particularly with respect to species delimitation for tropical and subtropical species. In some cases, re-ranking or combining subspecies and/or varieties may enable more accurate species delimitation.

<p><strong>A morpho-molecular re-appraisal of <em>Polystigma fulvum</em> and <em>P. rubrum </em>(<em>Polystigma</em>, Polystigmataceae)</strong></p>

Collections of eleven Prunus specimens infected with Polystigma species from Belarus and Russia yielded two existing taxa: Polystigma fulvum (sexual morph) and Polystigma rubrum (asexual morph). DNA based phylogenies of large subunit nuclear rDNA (LSU) and nuclear ribosomal internal transcribed spacer (ITS) are provided for the first time for Polystigma fulvum and its placement is confirmed within Polystigmataceae. The concatenated LSU and ITS DNA sequence data for P. rubrum, analyzed to infer its potential relationship to other Polystigma species, also show that strains of P. rubrum are segregated into two subclades with sufficient genetic differences. No significant differences in morphology or morphometry among the strains of P. rubrum between the two subclades, especially vis-à-vis the conidiogenous cell and conidium sizes are observed (P>0.05). Subspecies concept of P. rubrum complex is discussed.

Morphometric characteristics of Alpine salamanders: a support for subspecies validation and conservation?

Abstract The subspecies concept is not only useful to assess the evolutionary history of species and therefore describe their evolutionary potential, but it also has corollaries for defining conservation units and their management. Within Alpine salamanders, the subspecies status of Salamandra atra prenjensis, isolated in the Dinarides from its nominal subspecies Salamandra atra atra that occurs in the Alps, has been under debate. To remediate this fuzzy taxonomy, the present study investigates 14 morphological traits of Alpine salamanders originating from Austria and from Bosnia and Herzegovina (B&H). Multivariate analyses support a geographical structuring of morphological variation and the differentiation between the Dinaric (B&H) and Alpine (Austrian) populations. Within populations, a different correlation pattern among traits is registered, reflecting the distinct genetic architecture of multivariate phenotypes. This morphometric study supports recent molecular evidences of a strong differentiation between the Dinaric and Alpine populations and pleads in favour of the separate subspecies status, although a wider sampling of other populations and the inclusion of additional characters would be necessary to reinforce this conclusion. The recognition of Salamandra atra prenjensis as a distinct subspecies would highly contribute to the better conservation of this emblematic salamander.

Phylogeography and taxonomy of the barred grass snake (Natrix helvetica), with a discussion of the subspecies category in zoology

Using 12 microsatellite loci, mitochondrial DNA sequences and previously published morphological evidence, we examined the phylogeographic and taxonomic structure of Natrix helvetica. Our results support tentatively the recognition of five subspecies: (1) N. h. helvetica (Lacepède, 1789) from Western Europe, (2) N. h. cetti Gené, 1839 from Sardinia, (3) N. h. corsa (Hecht, 1930) from Corsica, (4) N. h. lanzai Kramer, 1970 from the Po drainage and peninsular Italy, except for parts of Calabria and perhaps Apulia, and (5) N. h. sicula (Cuvier, 1829) from Sicily and parts of Calabria. The status of the subspecies from Corsica and Sardinia warrants further research. Grass snakes from the two islands are genetically deeply divergent from their continental and Sicilian conspecifics. However, the mitochondrial haplotypes of N. h. cetti and N. h. corsa are not reciprocally monophyletic, and our analyses of microsatellite data did not unambiguously support their distinctness. Natrix helvetica lanzai comprises three mitochondrial lineages and could represent more than one taxon. Grass snakes of the northern mitochondrial lineage of N. h. lanzai (mainly from the Po drainage system) were previously assigned to N. h. helvetica. We propose that the recognition of subspecies within N. helvetica reflects its genetic differentiation best. Our subspecies concept resembles the definition of Evolutionarily Significant Units (ESUs) in that subspecies should be ideally confirmed by two independent genetic lines of evidence (mtDNA, nuclear genomic markers). In contrast to species, subspecies are still fully capable of extensive gene flow and may disappear in the evolutionary process as a consequence of secondary contact. Thus, subspecies represent an early stage of (incomplete) speciation. Using the subspecies category facilitates communication within and beyond science (legislation, conservation), a clear advantage compared to the cumbersome concept of ESUs.

The subspecies concept in Geocorinae: an integrated taxonomic case study on Geocoris (Piocoris) erythrocephalus (Lepeletier Serville, 1825) (Hemiptera: Heteroptera: Geocoridae).

The validity and necessity of subspecies as a taxonomic category and the implications of the subspecies concept in various taxa of animals is subject of debates since its very existence. In case of the species of the lygaeoid subfamily Geocorinae there are multiple examples of species consisting of up to nine subspecies which were mostly described as varieties or forms before the middle of 20th century, but upgraded to valid subspecies subsequently, usually without providing any arguments. As part of an integrated taxonomic study on the Palaearctic representatives of Geocorinae, the status of the subspecies of Geocoris (Piocoris) erythrocephalus (Lepeletier Serville, 1825) was revisited. A critical review of the literature available and our studies, involving analysis of COI sequences, morphological examination and distribution data processing with the use of Geographic Information System, concluded that two of the three subspecies, G. e. erythrocephalus and G. e. marginellus Horváth, 1907, can be considered valid, while G. e. litoreus Horváth, 1895, is merely a phenotypically manifested infrasubspecific genetic variety of G. e. erythrocephalus, with no taxonomic value as it was suggested by earlier study of another author. Besides the interpretation of evidence, the applicability of the subspecies concept in Geocorinae is discussed as well.

Integrative taxonomy improves delimitation in Hypericum subspecies

Integrative approaches have been very useful to identify diagnostic morphological characters in species delimitation and to understand how abiotic factors influence the geographical distribution of taxa and whether these factors may be driving to specific changes in phenotypic variation patterns. Here we use a South American Hypericum species to discuss the importance of such collaborations for taxonomy. The current classification recognizes four sympatric subspecies within Hypericum rigidum; however, due to the presence of intermediary diagnostic characters, a significant number of specimens cannot be assigned to any of these subspecies. Therefore, since the described subspecies are difficult to differentiate, in this study, we applied ecological and morphometric approaches to evaluate whether the infraspecific classification may be sustained in terms of environmental and morphological variation. Applied statistical analyses make evident the high morphological variation within H. rigidum subspecies and the absence of diagnostic characters that sustain the current infraspecific classification. Multivariate analyses also indicate that the morphologic variation of H. rigidum subspecies does not represent adaptive matching of phenotypes to the environment and, therefore, cannot be explained by phenotypic plasticity nor by ecotypic variation. We also discuss the non-standardization of the subspecies concept in plants. In conclusion, H. rigidum subspecies overlap geographically, ecologically and morphologically and, therefore, we recommend that these phenotypes should not be regarded as subspecies, but as part of one single highly variable species.

Arnica montana subsp. atlantica: Really a subspecies?

In Arnica montana L. (Asteraceae) two subspecies are described, A. montana subsp. atlantica (AMA), present only on the Iberian Peninsula and A. montana subsp. montana (AMM) with a very wide distribution area. The morphological differences between the two subspecies are small and variable. Therefore, this concept is sometimes questioned. To establish the genetic background of the two subspecies, populations of AMA and AMM together with herbarium samples and DNA Bank material of AMM were tested with 12 microsatellite markers. A. montana propagates by seeds or by clonal propagation of its rhizome. In AMA, clonality was frequent while in AMM only one case of clonality could be identified. Therefore, further results were clone-corrected. Genetically, AMA separated very well from AMM with a GST between the subspecies of 0.81, genetically justifying the subspecies concept of A. montana. Genetic variability in AMA (Hexp = 0.28) was lower than in the AMM populations (Hexp = 0.70). A somewhat higher fixation index of AMA (FST = 0.17, compared to an FST = 0.08 for AMM) may indicate that geneflow in AMA is a bit more restricted than in alpine AMM. However, the fixation index of AMA is not deviating from Hardy–Weinberg equilibrium. No inbreeding was observed for AMA (FIS = 0.10) and AMM (FIS = 0.08).

Subspecies definitions and legislation: from eastern wallaroo (Osphranter robustus robustus) to euro (Osphranter robustus erubescens)

As a consequence of genetic studies of population structuring, the usefulness of subspecies has been questioned, with opinions divided. The situation is further confused by the use of varying species and subspecies concepts. Most alternatives require each taxon to be following an independent evolutionary trajectory. These include traditional approaches and the more recent phylogenetic species concept. The latter has led to large increases in the apparent number of taxa in some groups, though strong objections have been raised to this approach. An alternative, the ecological species concept, has been opposed by phylogeneticists. These two approaches are compared using morphological and genetic data from common wallaroo (Osphranter robustus) populations. The different taxonomies that might result (many species, one species with two subspecies, one species with no subspecies) can have significant consequences for legislative and management decisions. The ecological approach is considered preferable for subspecies and the present taxonomy is maintained. A potential location of the boundary between the wallaroo subspecies is proposed. How the use of the different subspecies definitions would affect legislative decisions is explored.

The conundrum of subspecies: morphological diversity among desert populations of the California vole (Microtus californicus, Cricetidae)

Author(s): Patton, JL; Conroy, CJ | Abstract: We examined geographic trends in 4 morphological data sets (both craniodental and colorimetric measurements and scores of cranial foramina and qualitative craniodental variables) within and among 5 subspecies of the California vole, Microtus californicus, that occur within or adjacent to the Mojave Desert in eastern California. These analyses are corollary to those previously published on the same samples using both mitochondrial and nuclear gene sequences and microsatellite loci. The morphological and molecular data sets are generally concordant in supporting the existing infraspecific taxonomy, although important conflicts are evident. With the combination of data sets, we are able to assign previously unknown samples to these infraspecific taxa. We define and provide an emended diagnosis of each subspecies using the criteria established by Willi Hennig in his 1966 assessment of the qualities and characteristics of infraspecific taxa, which he understood to be fundamentally different than the hierarchical phylogenetic system he established as the conceptual framework for taxa at the species level and above. Hennig's views are contrary to how the subspecies concept has been applied in much of recent mammalian taxonomy.

Why we should develop guidelines and quantitative standards for using genetic data to delimit subspecies for data‐poor organisms like cetaceans

AbstractObtaining the representative morphological data required for traditional taxonomy is difficult, and sometimes impossible, for cetaceans, especially large ones. As a result, three quarters of the 88 currently recognized extant species have no subspecies and 40 taxa likely have additional unnamed taxa. Conservation needs give urgency to improving taxonomy because unnamed taxa are unlikely to receive protection equivalent to that received by named taxa. Genetic data can improve efforts to delimit subspecies, but the markers and methods used have varied and the magnitude of genetic difference used to justify subspecies distinctions across studies has also varied. Here, we define the concepts of populations, subspecies, and species to establish a foundation for developing guidelines (data to include and analyses to conduct) and quantitative standards (the magnitude of differentiation expected at different taxonomic levels) for using genetic data to support taxonomic recognition. Our definition is particularly applicable to data‐poor groups because it allows for naming a subspecies when there is uncertainty about whether lineages have diverged sufficiently for species‐level recognition. This allows a species that lacks convincing data for lineage divergence to be recognized as a subspecies while sufficient data are accrued, which could take decades for some cetaceans.

Ammonites, inoceramids and stable carbon isotopes of the Cenomanian–Turonian OAE2 interval in central Europe: Pecínov quarry, Bohemian Cretaceous Basin (Czech Republic)

In a well-exposed succession of Cenomanian through lower Turonian strata at Pecínov (Bohemian Cretaceous Basin, Czech Republic), a detailed study of ammonite and inoceramid assemblages was undertaken for the first time and combined with a revision of physical and carbon-isotope stratigraphy. The upper Cenomanian part of the OAE2 interval yielded a relatively rich ammonite assemblage containing Metoicoceras geslinianum (d'Orbigny, 1850), Calycoceras (Calycoceras) naviculare (Mantell, 1822), Pseudocalycoceras angolaense (Spath, 1931), Lotzeites aberrans (Kossmat, 1895), Euomphaloceras septemseriatum (Cragin, 1893), Sciponoceras gracile (Shumard, 1860), Allocrioceras annulatum (Shumard, 1860) and ?Puzosia sp. Inoceramid bivalves include Inoceramus pictus J. de C. Sowerby, 1829, Inoceramus sp. aff. Mytiloides praeturonicus Tröger, 2015 and Inoceramus cf. bohemicus Leonhard, 1897 in the Cenomanian part of the succession. The subspecies concept of the Inoceramus pictus group is modified here. The occurrence of Mytiloides puebloensis Walaszczyk and Cobban, 2000 indicates the lowermost Turonian Watinoceras devonense Zone. Earlier interpretation of the positive carbon-isotope excursion is refined here and a combined bio- and chemostratigraphic correlation is presented to successions from the Western Interior Seaway and the British Chalk.

USE OF PHYLOGENETIC ANALYSIS TO IDENTIFY EVOLUTIONARILY SIGNIFICANT UNITS FOR THE ORANGE-FRONTED PARAKEET (EUPSITTULA CANICULARIS) IN MEXICO

In avian conservation biology, the subspecies concept based on reciprocal monophyly has been successfully applied to define priority populations through Evolutionarily Significant Units (ESUs). In México, the Orange-fronted Parakeet (Eupsittula canicularis) ranks first in illegal parrot trade. Its distribution ranges from southern Sonora to Chiapas on the Pacific slope, with populations representing three subspecies: E. c. canicularis, E. c. eburnirostrum, and E. c. clarae. To identify and propose ESUs to assist in conservation proposals for different populations, we assessed subspecific reciprocal monophyly via phylogenetic analyses and haplotype networks based on the mitochondrial DNA genes cytochrome oxidase I y NADH dehydrogenase 2. Feather and blood samples from specimens collected from nests in 2005 and 2007 were used. A total of five specimens of E. c. eburnirostrum from two localities in the state of Michoacán and four specimens of E. c. clarae from the state of Sinaloa were analyzed and no specimens of E. c. canicularis were included. The analyses included sequences obtained by us and those previously reported for E. aurea, E. cactorum, E. canicularis, E. nana, and E. pertinax. Both the phylogenetic analyses and haplotype networks suggest two groups that correspond to two subspecies of E. canicularis based on morphological and geographical evidence. Therefore these two subspecies are proposed as independent ESUs for conservation purposes.

Clarifying Subspecies of Peregrine Falcons Along the Lost Coast of Alaska

Abstract The concept of subspecies is an important tool to help categorize and conserve biodiversity; thus, delineating the range of subspecies can have important management and conservation implications. The Peregrine Falcon (Falco peregrinus) is a widespread species that occurs throughout North America, where three subspecies are recognized: F. p. anatum, F. p. pealei, and F. p. tundrius. In Alaska, all three subspecies breed and their general distributions during the breeding season are well documented. However, the limits of their distributions were unclear or unconfirmed, especially those of F. p. anatum and F. p. pealei along the Lost Coast in the northeastern Gulf of Alaska. We describe plumage, morphology, and/or movements of Peregrine Falcons known to have nested (n = 6) or hatched (n = 3) within the Lost Coast and used this information to determine their subspecific group. For all nine birds, we found these characteristics to be consistent with F. p. anatum. Our results underscore the import...

The fate of subspecies category in Zoological systematics. 2. the present

The current approach to the use of the subspecies category in zoological systematics is an integrative one. This approach necessarily requires genetic confirmation of validity of the subspecies defined by morphological data. This makes it possible to separate the subspecies that really exist as separate monophyletic population groups, thereby clearing the system of many “phantom” taxa established in the course of noncritical use of the subspecies concept. However, a detailed analysis of intraspecific variation by molecular taxonomy methods in many cases reveals rather a complicated divergence pattern that cannot be adequately described in terms of the classic scheme of species and its subspecies. In light of the irregularity of intraspecific divergence rate with the involvement of molecular and morphological characters, it is proposed to use an extended system of subspecies taxa in the description of extra complicated situations. In addition to the subspecies, categories such as allospecies, morphotype, and morphospecies may be used; operational definitions are suggested for these categories. The systematics of the Palearctic great pond snails (the complex Lymnaea stagnalis s. lato) is used as a case study. The provisional system of this group developed by the author is based on morphological and phylogeographical data. The series of subspecies categories of different levels make it possible to maximally completely reflect the intraspecific variation of the great pond snails and, to some extent, the process of their genetic divergence and establishment of their distribution range.

Clines, Clusters, and Clades in the Race Debate

In this paper, I argue against Andreasen and Sesardic, who have both claimed that recent cluster results in population genetics serve as evidence that the human species contains, or at least once contained, subspecies. I show that the cluster results are in fact evidentially inert relative to each author’s preferred subspecies concept. I then sketch the kinds of biological facts that could be used to push the debate further.

Evidence for two subspecies of Gunnison’s prairie dogs (Cynomys gunnisoni), and the general importance of the subspecies concept

Accurate taxonomy is essential for conservation, but subspecies-level systematics can be hampered both by a lack of consensus on what constitutes a subspecies and by discordance among data types (e.g., genetics vs. morphology). Here we provide a framework for evaluating subspecies using multidimensional criteria, and suggest that taxa must satisfy multiple criteria to qualify as subspecies. As a case study, we use the Gunnison’s prairie dog (Cynomys gunnisoni), a species for which there has been disagreement regarding the existence of subspecies due to inconsistent application of criteria for defining subspecies. To explicitly test the hypothesis that two subspecies exist, we generated five predictions that could be evaluated with genetic data, while also using morphological and ecological criteria. We sampled 838 Gunnison’s prairie dogs from across the species range and performed a series of genetic analyses using 16 microsatellite and two mitochondrial loci (cytochrome b and the control region). We compared subspecies morphology and quantitatively evaluated whether abiotic and biotic habitat characteristics encountered by each subspecies differed. Genetic results from all five predictions supported the existence of two distinct subspecies within the confines of a proposed revision in the boundary between subspecies. The subspecies differed marginally in morphology and significantly in their habitats, suggesting ecological differentiation. Our results, which are in line with historical descriptions of morphologically distinct subspecies, suggest the subspecies should be recognized. This work provides support for the utility of integrating multiple data and analysis types to inform systematics and conservation.